Immunoassay for venom detection including noninvasive sample collection

• US 7,927,828 B2
• Filed: 10/17/2006
• Issued: 04/19/2011
• Est. Priority Date: 10/17/2005
• Status: Active Grant

• Current Assignee: Spidertech Inc., United States Of America As Represented By The Secretary Of The Air Force
• Original Assignee: Spidertech
• Inventors: McGlasson, David L., Stoecker, William V., Gomez, Hernan F., Green, Jonathan A.
• Primary Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US11/550,130
• Publication Number:
  US 20070141626A1
• Time in Patent Office: 1,645 Days
• Field of Search: 422/58, 422/61, 435/7.1, 435/970, 435/4, 435/287.1, 435/287.2, 435/287.7, 435/287.8, 435/975, 436/514, 436/518, 436/169, 436/170, 436/810, 436/808
• US Class Current: 435/7.92
• CPC Class Codes: G01N 2800/42 Poisoning, e.g. from bites or stings
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

Methods and immunoassays for diagnosing a bite or sting of a venomous organism in a patient having symptoms consistent with such a bite or sting are provided. A sample of venom is collected from the area of the suspected bite or sting using a swab and then contacted with an antibody that specifically binds to an antigenic site on venom present in the sample. Binding is then detected. The invention is illustrated by examples showing diagnosis of brown recluse spider bite, distinguishing it from other diagnoses with which it is often confused. This extremely sensitive test can detect venom antigens down to about 20 picograms even after the sample has been shipped and stored for periods of up to three weeks during the summer.

Claims

1. An immunoassay kit comprising:

(a) at least one antibody capable of binding to an antigen present in a Loxosceles venom;

(b) a swab for collecting a sample comprising venom from an area of a Loxosceles bite on a patient'"'"'s body; and

(c) a tracer for detecting binding between said antibody and said antigen;

wherein said immunoassay kit is capable of detecting Loxosceles venom present in a sample collected on said swab from the area of the Loxosceles bite.

2. The immunoassay kit of claim 1 wherein said antibody is immobilized on a solid substrate.

3. The immunoassay kit of claim 2 wherein said solid substrate is said swab.

4. The immunoassay kit of claim 1 wherein said swab comprises an absorbent or adsorbent material selected the group consisting of cloth, gauze, sponge, polymeric foam, cotton, rayon, Dacron, or nylon fibers, polyurethane foam, paper, and absorbent or adsorbent brushes.

5. The immunoassay kit of claim 1 wherein said swab is attached to a handle.

6. The immunoassay kit of claim 1 wherein said swab comprises natural or synthetic fibers.

7. The immunoassay kit of claim 1 wherein said swab comprises bristles.

8. The immunoassay kit of claim 1 wherein at least one antibody capable of binding to an antigen present in Loxosceles venom is a polyclonal antibody.

9. The immunoassay kit of claim 8 wherein said polyclonal antibody has been raised in a rabbit.

10. The immunoassay kit of claim 1 wherein said tracer comprises a monoclonal antibody capable of binding to said antigen present in Loxosceles venom.

11. The immunoassay kit of claim 1 comprising a portable colorimetric assay.

12. The immunoassay kit of claim 1 wherein at least one antibody capable of binding to an antigen present in Loxosceles venom is immobilized on an absorbent support.

13. The immunoassay kit of claim 1 also comprising a label attached to said tracer that indicates binding of said antibody to said antigen, wherein said label is selected from the group consisting of enzymes, radioactive isotopes, and color particles.

14. The immunoassay kit of claim 1 also comprising means for lysing cells present in said sample.

15. The immunoassay kit of claim 1 wherein said antibody capable of binding to an antigen present in Loxosceles venom is an antibody against a protein component of Loxosceles reclusa venom.

16. The immunoassay kit of claim 1 wherein said antibody capable of binding to an antigen present in Loxosceles venom is an antibody against sphingomyelinase.

17. The immunoassay kit of claim 1 also comprising instructions for use thereof.

18. The immunoassay kit of claim 13 also comprising a label comprising alkaline phosphatase.

19. The immunoassay kit of claim 13 also comprising a label comprising horseradish peroxidase.

20. The immunoassay kit of claim 18 also comprising a blocking agent.

21. The immunoassay kit of claim 20 wherein said blocking agent comprises nonfat milk solids.